| 2001 |
CARD11 (CARMA1) is a CARD-containing MAGUK family scaffolding protein whose CARD domain associates specifically with the CARD domain of BCL10, and when expressed in cells induces NF-κB activation and BCL10 phosphorylation. |
Co-immunoprecipitation, domain mapping, cell transfection NF-κB reporter assay |
The Journal of biological chemistry |
High |
11278692
|
| 2001 |
CARMA1 binds BCL10 via CARD-CARD interaction, induces translocation of BCL10 from cytoplasm to perinuclear structures, and stimulates BCL10 phosphorylation and NF-κB activation. |
Co-immunoprecipitation, confocal microscopy, overexpression in cells |
FEBS letters |
High |
11356195
|
| 2002 |
CARD11 mediates NF-κB activation selectively downstream of TCR/CD28 (but not TNFα or dsRNA), functions upstream of the IKK complex, and requires its CARD domain to cooperate with BCL10; CARD, coiled-coil, SH3, and GUK domains are each critical for signaling. |
Expression cloning, RNAi knockdown, RNAi-rescue domain mutagenesis, NF-κB reporter assay |
The EMBO journal |
High |
12356734
|
| 2002 |
CARMA1 is constitutively associated with lipid rafts; TCR stimulation induces physical association of CARMA1 with TCR and BCL10, and BCL10 translocates into lipid rafts upon TCR engagement. A CARMA1 mutant defective for BCL10 binding acts as dominant-negative for TCR-induced NF-κB and JNK activation. |
Co-immunoprecipitation, lipid raft fractionation, dominant-negative transfection, cytokine assay |
Nature immunology |
High |
12154360
|
| 2002 |
CARMA1 deficiency selectively impairs TCR-induced NF-κB activation and IL-2 production; reconstitution of CARMA1-deficient T cells with CARMA1 fully rescues the signaling defect. |
Somatic mutagenesis screen, reconstitution assay, NF-κB reporter, cytokine ELISA |
Nature immunology |
High |
12154356
|
| 2003 |
Genetic inactivation of CARD11 in mice causes a complete block in T and B cell immunity with a selective defect in JNK and NF-κB activation downstream of antigen receptors and PKC; CARD11 is also required for TLR4/LPS-induced B cell proliferation and JNK activation. |
Genetic knockout mouse, proliferation assay, cytokine production, kinase activation assays |
Immunity |
High |
12818158
|
| 2003 |
A point mutation (L232R equivalent) in the CARD11 coiled-coil domain abolishes antigen receptor-specific NF-κB and JNK signaling while preserving calcium/NFAT/ERK signaling, identifying the coiled-coil domain as essential for pathway-specific signaling. |
ENU forward genetic screen, reconstitution/complementation, signaling assays |
Immunity |
High |
12818157
|
| 2003 |
CARMA1 is required for recruitment of BCL10 to clustered TCR complexes and lipid rafts; its CARD domain is essential for engaging downstream signaling components in vivo. |
Knockout mouse, CARD-deleted knock-in mouse, lipid raft fractionation, NF-κB and proliferation assays |
Current biology : CB |
High |
12867037 12867038
|
| 2004 |
CARMA1 membrane association (via its MAGUK domain) is essential for function; CARMA1 recruits PKCθ, BCL10, and IKKβ into lipid rafts at the immunological synapse in a signal-dependent manner; BCL10-CARD fusion restores signaling in CARMA1-deficient cells lacking CARD. |
Reconstitution of CARMA1-deficient T cells, lipid raft fractionation, immunofluorescence, NF-κB reporter |
Molecular and cellular biology |
High |
14673152
|
| 2004 |
CARMA1 controls entry of IKK into lipid raft aggregates and the central region (cSMAC) of the immune synapse downstream of PKC, without affecting overall synapse organization or TCR/LFA-1/PKCθ recruitment to the synapse. |
Knockout T cells, peptide-specific imaging system, lipid raft fractionation, IKK activity assay |
The Journal of experimental medicine |
High |
15520247
|
| 2004 |
MALT1 is recruited to lipid rafts of the immunological synapse downstream of CARMA1 (CARMA1-deficient cells fail to recruit MALT1); MALT1 associates with CARMA1 in a BCL10-independent manner, and MALT1/BCL10/CARMA1 form a trimolecular complex. |
Lipid raft fractionation, Co-immunoprecipitation in CARMA1-deficient and reconstituted cells, dominant-negative deletion mutant assay |
The Journal of biological chemistry |
High |
14754896
|
| 2004 |
CARMA1 physically associates with IKKγ/NEMO and participates in an inducible large complex containing IKKγ/NEMO, BCL10, and IKKα/β; expression of the NEMO-binding region of CARMA3 exerts dominant-negative effects on BCL10-mediated NF-κB activation. |
Co-immunoprecipitation, dominant-negative overexpression, NF-κB reporter assay |
The Journal of biological chemistry |
Medium |
15184390
|
| 2005 |
BCR engagement induces progressive recruitment of CARMA1 into lipid rafts and association with PKCβ, which phosphorylates CARMA1 at S564, S649, and S657 in the serine-rich linker domain; mutation of S564 and S657 ablates CARMA1 function, whereas deletion of the linker causes constitutive, PKC-independent NF-κB activation. |
Lipid raft fractionation, in vitro kinase assay, site-directed mutagenesis, NF-κB reporter assay |
Immunity |
High |
16356855
|
| 2005 |
PKCθ phosphorylates CARMA1 in vitro and in vivo at Ser552 (linker region) following TCR-CD28 costimulation; CARMA1 S552A mutant fails to mediate TCR-induced NF-κB activation in CARMA1-deficient T cells and fails to recruit downstream signaling components to the immunological synapse. |
In vitro kinase assay, phospho-specific antibody, site-directed mutagenesis, reconstitution of CARMA1-deficient T cells, immunological synapse imaging |
Immunity |
High |
16356856
|
| 2005 |
TAK1 is required for IKK activation in BCR signaling and interacts with phosphorylated CARMA1; this interaction is mediated by PKCβ. IKK is also recruited to the CARMA1-BCL10-MALT1 complex in a PKCβ-dependent manner. |
Co-immunoprecipitation, kinase assay, siRNA knockdown, PKCβ-deficient B cells |
The Journal of experimental medicine |
High |
16301747
|
| 2006 |
IKKβ is required for initial CBM complex formation; upon engagement with the Carma1/BCL10/MALT1 complex, IKKβ phosphorylates BCL10 at its C-terminus, disrupting BCL10-MALT1 association and BCL10-mediated IKKγ ubiquitination — a negative regulatory feedback step. |
Genetic IKKβ-deficient T cells, in vitro kinase assay, mutagenesis of BCL10 phosphorylation sites, co-immunoprecipitation, cytokine measurement |
Molecular cell |
High |
16818229
|
| 2006 |
CaMKII is redistributed to the immune synapse upon T cell activation, phosphorylates CARMA1 on Ser109, and this phosphorylation facilitates the CARMA1-BCL10 interaction to enhance NF-κB activation. |
Computational expression analysis, CaMKII inhibitor/siRNA, in vitro kinase assay, Co-immunoprecipitation, NF-κB reporter |
Molecular and cellular biology |
Medium |
16809782
|
| 2006 |
CARMA1 selectively regulates JNK2 (not JNK1) downstream of TCR signaling; BCL10 is inducibly associated with JNK2 and acts as a JIP-like scaffold assembling JNK2, MKK7, and TAK1; this CARMA1/BCL10-JNK2 axis controls c-Jun protein levels. |
Genetic KO mice for CARMA1/BCL10, Co-immunoprecipitation, JNK isoform-specific kinase assay |
Immunity |
High |
17189706
|
| 2007 |
CARMA1 undergoes K48-linked polyubiquitination and proteasome-dependent degradation upon antigen receptor activation; ubiquitin acceptor sites reside in the SH3 and GUK domains; deletion of the Hook domain between SH3 and GUK induces constitutive ubiquitination; elimination of ubiquitination sites elevates NF-κB and JNK activation. |
Site-directed mutagenesis, proteasome inhibitor treatment, ubiquitin linkage-specific antibodies, NF-κB reporter, JNK assay |
Molecular and cellular biology |
High |
20008554
|
| 2007 |
IKKβ contributes to formation of the CARMA1-BCL10-MALT1 complex in B cells by phosphorylating CARMA1 (in addition to PKCβ-mediated phosphorylation at Ser668); PKCβ-mediated Ser668 phosphorylation is essential for BCR-induced CBM association and subsequent IKK activation. |
Phospho-specific antibodies, PKCβ-deficient B cells, IKKβ inhibitor, Co-immunoprecipitation, kinase assay |
The Journal of experimental medicine |
High |
18086859
|
| 2007 |
Caveolin-1 ligation of CD26 induces T cell NF-κB activation; the cytoplasmic tail of CD26 interacts with CARMA1, recruiting a complex of CD26/CARMA1/BCL10/IKK to lipid rafts. |
Co-immunoprecipitation, lipid raft fractionation, NF-κB reporter, soluble caveolin-1-Fc fusion protein stimulation |
The Journal of biological chemistry |
Medium |
17287217
|
| 2007 |
CARMA1 coiled-coil domain mediates constitutive oligomerization of CARMA1; disruption of coiled-coil structure impairs oligomerization and abrogates NF-κB activation; the CC1 subdomain is required for CARMA1 subcellular localization and CC2 for self-association. |
Co-immunoprecipitation of CARMA1 with itself, coiled-coil mutagenesis, subcellular localization imaging, NF-κB reporter |
The Journal of biological chemistry |
Medium |
17428801
|
| 2008 |
CARD11 contains a PKC-responsive inhibitory domain (ID) that controls the association of CARD11 with multiple signaling cofactors (BCL10, TRAF6, TAK1, IKKγ, caspase-8) through intramolecular interactions requiring both CARD and coiled-coil domains; TRAF6 and caspase-8 associate with CARD11 in a signal-inducible manner; CARD11 can recruit cofactors independently of BCL10 or MALT1. |
Domain mutagenesis, RNAi rescue assay, Co-immunoprecipitation in BCL10/MALT1-deficient cells, NF-κB reporter |
Molecular and cellular biology |
High |
18625728
|
| 2008 |
Oncogenic CARD11 coiled-coil domain mutations (e.g., L225LI) found in ABC-DLBCL induce constitutive NF-κB activation and enhance NF-κB activity upon antigen receptor stimulation when introduced into lymphoma cell lines. |
Sanger sequencing of DLBCL biopsies, transfection of mutant constructs into lymphoma cell lines, NF-κB reporter assay |
Science |
High |
18323416
|
| 2009 |
Cbl-b E3 ligase monoubiquitinates CARMA1, which disrupts CARMA1-BCL10 complex formation (without affecting CARMA1 protein stability), thereby suppressing NF-κB activation during NKT cell anergy. |
Co-immunoprecipitation, ubiquitination assay, Cbl-b KO mice, CARMA1 KO NKT cells, IFN-γ production |
Proceedings of the National Academy of Sciences of the United States of America |
High |
19815501
|
| 2009 |
PP2A (regulatory subunit Aα/PPP2R1A) interacts with CARMA1 and dephosphorylates PKCθ-dependent Ser645 phosphorylation on CARMA1; maintenance of this phosphorylation correlates with increased CBM complex formation, NF-κB activation, and IL-2/IFN-γ production. |
Co-immunoprecipitation, siRNA knockdown, in vitro dephosphorylation assay, phospho-specific antibodies, cytokine assay |
The EMBO journal |
High |
21157432
|
| 2009 |
HPK1 kinase interacts with CARMA1 in a TCR stimulation-dependent manner and phosphorylates the CARMA1 linker region at residues S549, S551, and S552 (distinct from PKC sites); CARMA1 S551A or S549A/S551A mutants fail to restore HPK1-mediated NF-κB activation in CARMA1-deficient T cells. |
Co-immunoprecipitation, in vitro kinase assay, site-directed mutagenesis, reconstitution of CARMA1-deficient cells, NF-κB reporter |
Proceedings of the National Academy of Sciences of the United States of America |
High |
19706536
|
| 2009 |
CARMA1 serine 649 phosphorylation, occurring later and more persistently than S657 phosphorylation after antigen receptor stimulation, down-regulates CARMA1 activity; S649A mutation promotes enhanced IKK and JNK activation at low stimulation doses. |
Phospho-specific antibodies, site-directed mutagenesis, PKCβ-deficient B cells, DT40 CARMA1-deficient reconstitution, IKK/JNK assays |
Journal of immunology |
High |
19917688
|
| 2010 |
Oncogenic CARD11 mutations F123I and L225LI induce hyperactivity by disrupting autoinhibition by the CARD11 inhibitory domain (ID): they disrupt ID-mediated intramolecular interactions, bypass the requirement for ID phosphorylation during TCR signaling, and selectively enhance CARD11 affinity for BCL10. |
Co-immunoprecipitation, mutagenesis, dominant-negative ID peptide, phosphorylation-mimetic/defective mutants, NF-κB reporter |
Biochemistry |
High |
20799731
|
| 2010 |
Calcineurin (activated by calcium) positively regulates CBM complex formation by dephosphorylating BCL10, promoting CARMA1-BCL10-MALT1 assembly; calcineurin A is physically associated with the CBM complex. |
Cyclosporin A/FK506 pharmacological inhibition, EGTA-AM calcium chelation, calcineurin siRNA knockdown, in vitro dephosphorylation, Co-immunoprecipitation, NF-κB reporter |
The Journal of biological chemistry |
High |
21199863
|
| 2011 |
OX40 (a TNFR family member) assembles a signalosome containing TRAF2, RIP, IKKα/β/γ as well as CARMA1, MALT1, BCL10, and PKC in membrane microdomains independently of TCR engagement; NF-κB activation by OX40 requires CARMA1 and PKC. |
Co-immunoprecipitation, membrane microdomain fractionation, genetic-deficient cells, NF-κB reporter, T cell survival assay |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
21282629
|
| 2011 |
ADAP associates with CARMA1 and is required for TCR-stimulated assembly of the CARMA1-BCL10-MALT1 complex; a distinct region of ADAP mediates CARMA1 association and NF-κB activation, separable from ADAP's role in adhesion regulation. |
ADAP-deficient T cells, Co-immunoprecipitation, NF-κB nuclear translocation assay, IκBα phosphorylation/degradation |
Science |
High |
17478723
|
| 2013 |
The reconstituted CBM signalosome forms a helical filamentous assembly in which substoichiometric CARMA1 nucleates BCL10 CARD filaments in a highly cooperative process; crystallography, NMR, and EM reveal the Bcl10 CARD filament structure and CARMA1-BCL10 interface; structure-guided mutagenesis confirmed these interfaces for MALT1 activation in vitro and NF-κB activation in cells. |
In vitro reconstitution, cryo-EM, X-ray crystallography, NMR, electron microscopy, site-directed mutagenesis, NF-κB reporter |
Molecular cell |
High |
24074955
|
| 2013 |
USP9X deubiquitinating enzyme interacts with BCL10 in the CBM complex and removes TCR-induced ubiquitin chains from BCL10, which facilitates CARMA1-BCL10-MALT1 association and is required for NF-κB activation in T cells. |
Co-immunoprecipitation, USP9X siRNA knockdown, ubiquitination assay, NF-κB reporter, T cell proliferation/cytokine assay |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
23690623
|
| 2013 |
STUB1 (an E3 ubiquitin ligase) constitutively interacts with CARMA1 and catalyzes Lys-27-linked ubiquitination of CARMA1 upon TCR stimulation; STUB1 knockdown diminishes TCR-induced NF-κB activation and IL-2 production. |
Co-immunoprecipitation, siRNA knockdown, ubiquitination assay with linkage-specific mutant ubiquitin, NF-κB reporter, cytokine assay |
European journal of immunology |
Medium |
23322406
|
| 2014 |
CARMA1 (and MALT1 but not BCL10) is required for optimal TCR/CD28-induced mTORC1 activation in T cells; MALT1 catalytic activity is required for CD4+ T cell proliferation and increased metabolic flux in activated T cells. |
Genetic-deficient cells, MALT1 catalytic inhibitor, mTOR substrate phosphorylation assay, metabolic flux assay, T cell proliferation |
Science signaling |
High |
24917592
|
| 2015 |
CARMA1 clustering through SH3-GUK domain interactions (intra- and intermolecular) is required for activation-induced microcluster formation at the immunological synapse and for TCR-induced and oncogenic NF-κB signaling; disruption of SH3-GUK interactions abolishes CARMA1 microclusters and NF-κB activation. |
SH3/GUK mutagenesis, live-cell confocal imaging of synapse microclusters, Co-immunoprecipitation, NF-κB reporter, ABC-DLBCL cell lines |
Nature communications |
High |
25602919
|
| 2015 |
RNF181, an E3 ubiquitin ligase identified by BRIC screening, interacts with CARD11 and ubiquitinates BCL10 to negatively regulate antigen receptor signaling to NF-κB; RNF181 limits DLBCL cell proliferation dependent on aberrant CARD11 signaling. |
BRIC interaction screen, Co-immunoprecipitation, in vitro ubiquitination assay, BCL10 protein level assay, NF-κB reporter, DLBCL cell proliferation assay |
Molecular and cellular biology |
Medium |
26711259
|
| 2016 |
BCL10 undergoes TCR-induced linear (LUBAC/HOIP-mediated) polyubiquitination, which is required for association with NEMO/IKKγ; CARD11 promotes this by co-recruiting BCL10 and HOIP via its coiled-coil domain; oncogenic CARD11 variants spontaneously induce Lin(Ub)n-BCL10 correlating with enhanced HOIP and BCL10 binding. |
Linear ubiquitin-specific antibodies, site-directed mutagenesis of BCL10 lysines, HOIP/CARD11 domain mutagenesis, Co-immunoprecipitation, NF-κB reporter |
The Journal of biological chemistry |
High |
27777308
|
| 2016 |
The CARD11 inhibitory domain (ID) contains four repressive elements (REs) that function cooperatively and redundantly to prevent spontaneous NF-κB activation; each RE contributes to closed inactive state maintenance and prevention of BCL10 binding; oncogenic mutations disrupt intramolecular interactions mediated by multiple REs. |
Domain mutagenesis, intramolecular interaction assay, quantitative NF-κB reporter assay, domain-deletion analysis |
The Journal of biological chemistry |
High |
26884334 26884335
|
| 2017 |
Germline hypomorphic CARD11 mutations cause loss-of-function and dominant-interfering activity for antigen receptor-induced NF-κB and mTORC1 signaling; glutamine supplementation partially rescues mTORC1 and IFN-γ production defects, indicating CARD11 is required for glutamine import into T cells. |
Transfection of mutant constructs, NF-κB and mTORC1 reporter assays, patient primary T cell functional assays, glutamine supplementation rescue experiment |
Nature genetics |
High |
28628108
|
| 2018 |
Cryo-EM structure of BCL10 CARD filament at 4.0 Å redefines CARD-CARD interactions; BCL10 polymerizes in a unidirectional manner in time-lapse confocal imaging; CARMA1 serves as a hub nucleating star-shaped BCL10 filament networks and decreases the lag period of BCL10 polymerization; MALT1 immediately dimerizes on BCL10 filaments and TRAF6 cooperatively decorates CBM filaments in higher-order assemblies. |
Cryo-EM structure determination, time-lapse confocal imaging, in vitro polymerization assay, EM of MALT1/TRAF6 decoration |
Proceedings of the National Academy of Sciences of the United States of America |
High |
29382759
|
| 2019 |
Structure of a CARD9 region containing an extensive CARD-coiled-coil interface reveals the structural basis for autoinhibition; disruption of this interface leads to hyperactivation and Bcl10-templating filament formation in vitro for both CARD9 and CARD11; structure of the CARD9 filament illuminates mechanisms of CARD11 oncogenic mutations and how autoinhibition is relieved during canonical activation. |
X-ray crystallography of autoinhibited CARD9, cryo-EM of CARD9 filament, cell-based hyperactivation assays, in vitro filament polymerization |
Nature communications |
High |
31296852
|
| 2010 |
GAKIN (a kinesin-3 family member) associates with CARD11 in a signal-dependent manner, competes with BCL10 for CARD11 association, and dynamically localizes to the immunological synapse to redistribute CARD11 from the central to a distal region, thereby negatively regulating TCR signaling to NF-κB. |
Expression-cloning screen, Co-immunoprecipitation, live-cell imaging at immunological synapse, NF-κB reporter, competition binding assay |
Molecular cell |
High |
21145487
|
| 2012 |
A quantitative signaling screen identifies gain-of-function CARD11 mutations in the CARD and LATCH domains; LATCH domain functions with the CARD to promote autoinhibition; mutations in LATCH or CARD disrupt inhibitory domain binding, promote BCL10 association, induce BCL10 ubiquitination, NF-κB activation, and human lymphoma cell survival. |
High-throughput quantitative NF-κB signaling screen, domain mutagenesis, Co-immunoprecipitation, ubiquitination assay, lymphoma cell survival assay |
Molecular and cellular biology |
High |
23149938
|
| 2021 |
Complete human CARD11 deficiency (biallelic p.Arg837* loss-of-function) prevents CBM complex formation, severely impairs NF-κB, JNK, and MALT1 paracaspase activation in B and T cells, and causes a developmental block in B cells at the naive/type 1 transitional stage with impaired T follicular helper cell development. |
Whole-exome sequencing, immunoblot signaling assays, flow cytometry immunophenotyping, MALT1 paracaspase activity assay, transcriptomics, hematopoietic stem cell transplantation functional restoration |
The Journal of allergy and clinical immunology |
High |
33872653
|